JPH01242527A - Anti-cancer composition - Google Patents

Abstract

PURPOSE: To obtain an anticancer composition effective for treating melanoma and cancer cells and capable of selectively attacking cancer cells by compounding a specific cyanine compound. CONSTITUTION: This composition is obtained by compounding a cyanine compound of formula I [X1 and X2 are each O, S or Se; Z is an atom necessary for completing a 6-membered saturated carbon ring condensed with aromatic ring; A is CHCH, a group of formula II (R5 is methyl, ethyl, etc.), etc.; (n)=0, 1; R1 and R2 are each methyl or ethyl; R3 and R4 are each H or a halogen; Q is an anion]. As the compound of formula I, 3-methyl-2-[3-(3-methyl-4- benzothiazolin-1-ylidene-2-propen-3yl]benzothiazolium iodide, etc., is used. A compound of formula I is characterized in that it can highly selectively treat welldifferentiated carcinoma and melanoma of a mammalian by intravenous or peritoneal administration.

Description

DETAILED DESCRIPTION OF THE INVENTION [Industrial Field of Application] The present invention relates to an anticancer composition containing a cyanine compound.

[Conventional technology]

There is no need to repeat that cancer is a scourge in the modern world, especially in developed countries. According to estimates, out of 100,000 people in developed countries, 20.00
It is estimated that up to 0 people will develop cancer and will not receive effective treatment. rDynamicsand opp
ortunities in CancerManag
elementJ (SRI International)
(1985), Vol. 1, p. 6,
Approximately 5 million people are expected to die from cancer in 1986. This is serious because of the pain caused by cancer and the sense of helplessness that comes with actually dying.

It is therefore not surprising that many attempts have been made to discover anti-tumor agents.

The need for effective anti-tumor drugs is so well known that four news organizations request information whenever there is a rumor that one has been discovered.

One problem with conventional chemotherapy drugs in cancer treatment is that they tend to be highly toxic not only to cancer cells but also to healthy cells. As a result, said medicines may cause undesirable side effects such as alopecia, nausea, nephrotoxicity, precardiac disease (
cardiotoxicity). Another problem is that success rates are relatively low, even with the most reputable medications. For example, rAnt
Ibiotics Chemother J Vol.
24. As reported in p. 149-159 (197B), the well-known chemotherapeutic agent "Adriamycin" (trade name) has been used in the treatment of gastric cancer in Japan for 14 years.
．． It only showed an effective response rate of 5%.

As the problem of cancer remains serious, people take such medicines and suffer side effects as described above in the hope that the cancer will be alleviated before the side effects become intolerable. Although some selectivity with respect to toxicity is now available, i.e., chemotherapeutic agents can selectively kill cancer cells rather than healthy cells, many such conventional chemotherapeutic agents In this case, the selectivity does not exceed 2 or 3, for example, as defined by the IC3° value in human cancer in vitro studies.

This usual selectivity of 2 or 3 is not sufficient. This is because undesirable side effects still plague patients. For selectivity to be significant enough to predict a reduction in undesirable side effects, the IC5o
The selectivity, defined as a ratio, must be at least 5:1.

[Means to solve the problem]

Therefore, chemotherapeutic drugs with significant selective toxicity against at least some cancers have been a long-standing and needed problem to be solved. In particular, this request
differentiated cancer, as described in Cover, March 1986, page 37.
arcinoma) has been present in the field of differentiated cancer since it accounts for about 85% of cancers. Such cancers include so-called hollow J organs such as the brain, colon,
Includes cancers of the bladder, lung, prostate, stomach and pancreas.

There is a need for effective chemotherapeutic agents for colon cancer, as there are currently no known highly effective drugs for the disease (Discover, March 1986).
pages 36-46, especially page 38).

It has been previously suggested that cyanine dyes are useful in steaming processes. According to JP-A-54-157839, various cyanine pigments (actually dyes) are shown to be inhibitors of all types of cancer. This expansive claim is based solely on tests showing that the cyanines under test significantly inhibit the internal respiration of cancer cells (thus, this means that the cancer cells are dead). .

However, the dye's I
C. No results of clonogenic testing of the cyanines to establish values have been reported. There are also no in vivo data establishing whether any of the cyanines tested actually inhibit specific xenograft human tumors in nude mice.

After extensive screening and animal model studies, the present inventors have determined that there are other types of cyanine dyes that are more active against cancer than the cyanine dyes described in the above-mentioned Japanese Patent Publication. I found it. In fact, this group of cyanines exhibits unexpectedly remarkable selectivity for cancer cells, to the extent that they exclude normal cells, compared to the selectivity of the cyanines described in the above-mentioned Japanese Patent Application Publication No. . Such selectivity makes it possible for the first time in cancer chemotherapy to use a drug whose side effects are rarely debilitating to the disease itself.
It means that it has been confirmed.

[Means to solve the problem]

According to the present invention, the structural formula (wherein X and atom, A is =CH-CH= or R3, provided that if Xt is 0, A is the latter group, and n is O or 1, and R1, R1 and R1 are each independently a methyl group or an ethyl group, R
3 and R4 are each independently a hydrogen atom or a halogen atom, and Q is a pharmaceutically acceptable anion. Anti-cancer compositions are provided that are particularly effective for treating cells.

The compound of formula N) is known as a cyanine dye. Therefore, hereinafter, it may also be referred to as a cyanine dye.

The present invention will be explained in detail below.

As used herein, "differentiated cancer"
edcarcinomas) J include adenocarcinomas (this type includes about 90% of prostate cancers that occur in about 50% of men in the United States over the age of 65), most squamous cell carcinomas, and all transitional cell carcinomas. Includes cancer. The differentiated cancers mentioned above do not include oat cell carcinoma, large cell carcinoma in the lungs, and poorly differentiated cancers. These cancers are not considered to be affected by the present invention.

Effective treatment of differentiated cancers includes promoting tumor regression and mitigation;
Includes tumor regression and growth inhibition.

Thus, the types of cancers that can be treated by the compositions of the invention include all organs such as lungs (other than those mentioned above), colon, brain, bladder, prostate, pancreas, stomach, vagina, esophagus,
Includes cancers from the tongue, nasopharynx, liver, ovary and suisumaru.

The present invention is characterized by the use of a cyan dye represented by the above structural formula (1). Useful dyes within this group include:

3-Methyl-2-C3-(3-methyl-4-benzothiazolin-2-ylidene-2-propen-3-yl)penzothiazolium iodide; 3-ethyl-2-(3-(3
-ethyl-4-benzothiapurin-2-ylidene)-2
-propen-3-yl]-benzothiazolium iodide; 5-chloro-3=ethyl-2-(3-(3-ethyl-
4-Benzothiazolin-2-ylidene)-2-propen-3-yl]penzoselenazolium iodide; 3-ethyl-2-(3-ethyl-5-(2-(3-ethyl-4-
Benzoxazolin-2-ylidene)ethylidene)-4-
Oxo-2-thiazolidinylidenemethyl l-4,5,6
, 7-titrahydropenzothiazolium iodide; 5-
Chloro-3-methyl-2-(3-(3-methyl-4-benzothiazolin-2-ylidene)-2-propene-3-
yl]penzothiazolium iodide; 5-chloro-3-
Ethyl-2-(3-ethyl-5-C2-(3-ethyl-
4-benzoxapurin-2-ylidene)ethylidene 4-oxo-2-thiazolidineylidenemethyl l-4,
5,6,7-titrahydropenzothiazolium iodide; 3-methyl-2-(3-methyl-4-benzothiazolin-2-ylidenemethyl)penzoselenazolium iodide; 5-chloro-3- Methyl-2-(3-methyl-4=
benzothiazolin-2-ylidenemethyl)penzothiazolium iodide and 5-chloro-3-ethyl-2-(3
-ethyl-4-penzoxasoline=2-ylidenemethyl)-4,5,6,7-titrahydropenzothiazolium iodide. The above-mentioned compounds containing any of the counter ions described below in place of iosito are also included.

Particularly preferable ones are as follows.

3-Methyl-2-(3-(3-methyl-4-hendithiazolin-2-ylidene-2-propen-3-yl)pendthiazoline chloride; 3-ethyl-2-(3-(3
-ethyl-4-benzothiazolin-2-ylidene)-2
-propen-3-yl] pendzthiazoline iodide; 5
-chloro-3-ethyl-2-(3-(3-ethyl-4-
Benzothiazolin-2-ylidene)-2-propene-3
-yl] penzoselenazolium iodide; and 3-ethyl-2-(3-ethyl-5-(2-(3-ethyl-4-
Benzoxazolin-2-ylidene)ethylidene-4-oxo-2-thiazolidinylidenemethyl)-4,5,6,
7-titrahydropenzothiazolium iodide.

Any pharmaceutically acceptable anion,
Can be used with the cyanines of the invention. As used herein, "pharmaceutically acceptable anion" refers to an anion that is non-toxic to the host mammal and provides an aqueous solubility of the dye of at least 0.1 /a/. Examples include halides such as chloride, bromide, iosito, and fluoride; sulfonates such as aliphatic and aromatic sulfonates such as methanesulfonate, trifluoromethanesulfonate, p-toluenesulfonate (
tosylate), naphthalene sulfonate and 2-hydroxyethane sulfonate; sulfamates such as cyclohexane sulfamate; sulfates such as methyl sulfate and ethyl sulfate; bisulfates; porate; alkyl and dialkyl phosphates such as dimethyl phosphate, diethyl phosphate, and methyl hydrogen phosphate. ; pyrophosphates such as trimethyl pyrophosphate and diethyl hydrogen pyrophosphate; and carboxylates, preferably carboxy and hydroxy substituted carboxylates. Examples of such carboxylates include acetate, propionate, valerate, citrate, maleate, fumarate, lactate, succinate, tartrate and Henschede.

The most preferred anion is halide, since halide confers greater water solubility to the dye than other anions.

Both the cyanines of the present invention and the method for producing them are known per se.

Methods of administering the dye include intraperitoneal (IP), intravenous (IV) or intravesical infusion using a pharmaceutically acceptable carrier solvent or an imbedded drug pump. The IP administration method described above is particularly useful in treating cancer that has metastasized to the peritoneal cavity. For example, treatment of ovarian cancer is carried out by RP injection.

The pharmaceutically acceptable carrier can be any carrier, such as a solvent that sufficiently dissolves the cyanine dye. Examples of carrier solvents suitable for human use include 5% dextrose in water, or mixtures of polyols such as polyethoxymolized castor oil and ethanol (National C
“Diluent” from ancer In5titude
11h12'']. Additional acceptable carrier solvents include isotonic salt solutions for TV and IP injections, dimethyl sulfoxide (DMSO) for intravesical injections,
And in some cases it's just water.

However, fresh water is currently not preferred.

Other useful carriers include:

Ingredients such as gelatin, natural sugars such as sucrose or lactose, lecithin, pectin, starch (e.g. cornstarch), alginic acid, tylos
e), talc, lycobodium, silica (e.g. colloidal silica), glucose, cellulose, cellulose derivatives such as cellulose ethers (e.g. methyl hydroxypropyl cellulose, methyl cellulose, hydroxyethyl cellulose), stearates such as methyl stearate and glyceryl stearate, 1 carbon atom
2 to 22 fatty acids, especially magnesium and calcium salts of saturated acids (e.g. calcium stearate, calcium laurate, magnesium oleate, calcium palmitate, calcium behenate and magnesium stearate), emulsifiers, oils and fats, especially those of vegetable origin. (e.g. peanut oil, castor oil, olive oil, sesame oil, cottonseed oil, corn oil, malt oil, sunflower seed oil, cod liver oil), saturated fatty acids (C, □11□40□ to Cl81
1360□ and mixtures thereof) such as mono-, di- and triglycerides such as glyceryl monostearate, glyceryl distearate, glyceryl tristearate, glyceryl trilaurate, pharmaceutically compatible mono- or polyhydric alcohols and polyglycols. Evacricoline, mannitol, sorbitol, pentaerythritol, ethyl alcohol, diethylene glycol, triethylene glycol, ethylene glycol, propylene glycol, dipropylene glycol, polyethylene glycol 4
00, and other polyethylene glycols, and mN of these alcohols and polyglycols, saturated and unsaturated fatty acids (2 to 22 carbon atoms, especially 10 to 18 carbon atoms)
and monohydric aliphatic alcohols (alkanols having 1 to 20 carbon atoms) or polyhydric alcohols such as glycols, glycerin, diethylene glycol, pentaerythritol, sorbitol, mannitol, ethyl alcohol,
Esters with butyl alcohol, octadecyl alcohol etc. e.g. glyceryl stearate, glyceryl palmitate, glycol distearate, dilauryl & glycol, glycol diacetate, monoacetin, triacetin, glyceryl oleate, ethylene glycol stearate, etherified polyhydric alcohols ester of,
Benzyl benzoate, dioxolane, glycerin formal, tetrahydrofurfuryl alcohol, CI-%-C
Polyglycol ethers of I2 alcohols, dimethylacetamine, lactamide, lactates such as ethyl lactate, ethyl carbonate, silicones (particularly medium viscosity dimethylpolysiloxane), magnesium carbonate and the like.

Other additives and methods of preparing the compositions are described in the literature, for example in US Pat. No. 4,598,091.

The compositions of the present invention contain a therapeutically effective amount of the cyanine compound of 2N).

Based on the results obtained from the animal models described below, for human treatment, the dosage of the active cyanine component should be 50 to 100 μ/day in IV infusions every 3 weeks, 0.0% per day for 2 to 6 days. .5-1 dawn/kg, or 3 days or 6
It is estimated that it is 20 to 30 cm/day per week. Here, r/m J means, as usual, the surface area of the patient's body. This estimate is Go
odman &Gilman's r ThePharm
acological Ba5is of Thera
peutics J (6th edition) No. 1675-173
MacMillanPubli in New York State on page 7
shing Company, 1980).

[For production]

As mentioned above, one of the advantages of the present invention is that the cyanines selectively attack cancer cells rather than normal cells. Machine knowledge is not essential to the practice of the invention, but is believed to occur through preferential inhibition of oxygen consumption in the mitochondria of cancer cells. This selectivity is achieved by colonic
nic) further shown in in vitro studies. That is, C
Cell cultures of X-1 (human colon cancer) and CV-1 (normal monkey epithelial cells) were prepared by conventional methods and treated with various concentrations, and the concentration of dye that inhibited cell colony formation by 50% was determined by IC. ,. shall be. This IC. The values are CX-1 series and c
Compare with both of the vi series. That is, the value of CV-1 is divided by the value of CX-1. The value is lOn/rrJ for cv-i and 111g/r for CX-1.
d, the dye is 10 times more effective in killing colon cancer cells compared to monkey epithelial cells. This is expressed as a selectivity "S" of 10. In other words, to inhibit 50% of normal cells, 10 times the concentration of the drug is required to inhibit 50% of cancer cells, suggesting that the drug selectively kills cancer cells. are doing.

The selectivity and CX-1 values obtained by testing the four cyanines of the invention prepared under normal light conditions in the laboratory are shown in Table I below, along with a fifth cyanine for comparison purposes.

Kinpaku Tage [Examples] In the following examples, "nude mouse protocol" refers to conventional xenograft human cancer cells or melanoma cells transplanted into athymic mice. It is well established that efficacy against human cancers is estimated by testing in nude mouse-human xenograft models. This is, for example, Journal of the Nation
nal Cancer Institute, Vol.
, 74. t'h4, pages 899-903, especially page 889 (
April 1985).

The cyanines tested in animal model studies are:

The following margins are 112 indentations.'' LOX Melanoma Test Using Nude Mice The human melanoma cell line LOX, which grows rapidly when inoculated into the peritoneal cavity of athymic mice, was used.

N lesions subcutaneously inoculated into athymic mice were excised under sterile conditions and converted into single cell suspensions using a metal grid in a 4 ml mesh. The red blood cells were then incubated at 0°C for 20 min at 4°C.
．． Hemolysis was achieved by incubation with 17M ammonium chloride. Next, the viability of the cells was evaluated using trypan blue, and 5 million viable cells were placed in Du
lbecco modified Eagles medium (ohg) 0.1m
- group of athymic Swiss nu/nu mice (
Injected intraperitoneally on day 0). These mice were then randomly divided into treatment and control groups.

Treatment by intraperitoneal injection started the next day (1 day later). Mice receiving dye N11l or dye 11h2 received 1 kg/kg per day for 5 days, followed by 100 kg/day on the 8th day.
Day 122nd Day 144th and Day 166th 1.5 mg
/ kg, and thereafter 2 N/kg twice a week. One mouse each was used for the treatment and control groups. The control group is pigment! A 5% dextrose solution was given in the same volume as that of the lhl-treated group.

In the same experiment, the dye of Comparative Example 1 was administered to 10 mice according to the following procedure. 0.2■/kg per day for 5 days
, followed by 8th day, 100th day, 122nd day, 144th day and 1
0.5 ■/kg on day 66 and 1 ■/kg twice a week thereafter (low doses of this dye were given due to its toxicity; in this Comparative Example 1 dye 11h
In a preliminary study using a dose of 1, the mice died before reaching the 50% survival point of the controls, indicating that a lower dose was needed).

The purpose of this experiment was to
This shows the importance of the length of 2. That is, Comparative Example 1 differs from Example 2 in that R1 and R2 are propyl groups instead of ethyl groups.

This compound is the one described in Example 3 of JP-A-54-157839.

The test results are shown in Table H. T/C (%) is the treatment (t
The mean survival age of the rea Led) group and the untreated control (co
This is the ratio to the average survival age of the ntrol) group.

Table-E Example Dye IVhT/C I 1 205%2
2 176%3 3
168%4 4 16
6% Comparative Example 1 5 128% The results of Examples 1 and 2 are shown in FIGS. 1 and 2, and the results of Comparative Example 1 are shown in FIG. 3. In Figures 1 to 8, "Survival %
” is the percentage of the total group (treated group “T” or control group “C”) that is still alive on the day in question. The 50% data point was selected for the T/C calculation because it is a statistically significant point]. Any T/C value greater than 145% is generally considered by those skilled in the art to be an increase in survival or tumor control. Therefore, Examples 1-4 all demonstrate successful treatment of melanoma. However, in Comparative Example 1, R3 and R in the structure of formula (1)
It was not successful because 2 is a propyl group rather than an ethyl or methyl group. Although the reason for this is not fully understood, it is believed that it is partly due to stereospecific effects that occur when R1 and R2 are too long.

, LL2'-2 Ovarian cancer test using nude mouse protocol Dye N[11 and test of 2 are repeated, except that the cancer to be treated is 0VCAR-3, and 10 million human ovarian cancer cell lines of this was injected IP into athymic Swiss nu/nu mice. In Example 5, ingestion of Dye 1 by IP injection was started the day after cancer injection into 10 mice. 1.5■/kgzVt per minute for 4 days, 7th day, 10th day, 1
2 ry on days 3, 16, 19, and 21
g/kg, and thereafter twice a week, 2■/
kg. Ten mice with cancer injections as described above were selected as a control group and fed with 5% dextrose equal to the volume of the treated group.

In Example 6, 10 mice started receiving dye Yang 2 by IP injection the day after the cancer was injected. 1.5 kg/kg daily for 4 days, then 2 kg/kg on days 7, 10, 13, 16, 19, and 21, and twice a week thereafter. , 2■/kg. The same control group as in Example 5 was used. In Comparative Example 2, the test of Comparative Example 1 was repeated, but this time on ovarian cancer xenografts. Administer the dye at a rate of 0.2 seconds per minute for 4 days.
/kg, followed by 0.5■/kg on days 7, 10, 13, 16, 19 and 21, and then 0.5■ twice a week thereafter. /kg was used.

The results are shown in Table 3 and Figures 4 and 5.

5 1 238%6
2 157% Comparative Example 2 5
138% l21: Treatment of Mouse Bladder Cancer Using Intraperitoneally Administered Cyanine Dyes MB49 tumor cells (i.e., mouse cancer cells induced with oncogenic 7,12-dimethylbenzo(a)anthracene) were treated with normal male BDPI. Mice were injected intraperitoneally (2×10′ per mouse on day 0). Cyanine dye ml or N1
No. 12 was administered according to the following procedure. 0.2 erg on the first day
l kg, 0.4trg1kg on the second day, 0 on the third day
．． 8■/kg, 1.2■/kg on the 4th day, and 1.5■ on the 8th, 12th, 16th, 20th, and 24th days.
/kg. No subsequent injections were performed.

For pigment 5, 0.1 mg/kg on the first day;
0.2■/kg% on the second day, 0.3■/kg% on the third day
On the 4th day, it was 0.4■/kg, and on the 8th, 12th, 16th, 20th, and 24th days, it was 0.5■/kg. No subsequent injections were performed. Ten control mice were treated with the same volume of 5% dextrose as those treated with Dye Amount 1. Survival rates were calculated as in the previous example. The results are shown in Table ■.

Leave - 1 ■ 7 1 250%
8 2 190%
Comparative Example 3 5 113% These examples are shown in FIGS. 6 to 8, respectively. As with the other cancers treated, dye Yang 5 (similar to dyes A1 and A2, but with a propyl group) was not successful.

Comparison M The following cyanines were determined to be unacceptable because they had selectivity "S" values of less than 60 in the in vitro test described above. Its structure is expressed by the following formula.

R+ Ih Table 3 shows the selected groups and selectivity "S" values.

Comparison fi -shi--also L Z” 6
A-40S Benzo-C11
-Cl33C(Clls)z
Cyclohexano ・CH-CH8 Eel S
Se benzo io s
s benzo -CH-CH★
Nude mouse experiments were conducted on LOXi chromoma, and T/C
Mackerel Please compare with dye Nα3 in Table 1.

H, CI + 3
3.31=90% of the richness was never obtained.

〔Effect of the invention〕

An advantageous feature of the invention is that intravenous or intraperitoneal administration can be used to treat differentiated cancers and melanomas in mammals with high selectivity.

Another advantage of the present invention is that it performs the above treatment while minimizing undesirable side effects.

Yet another advantage of the present invention is that it provides an anti-cancer drug that is effective against colon cancer, a cancer that has traditionally been relatively immune to chemotherapy treatments.

[Brief explanation of the drawing]

Figures 1, 2, and 4-7 are graphs plotting cancer survival rates in animal models treated or not treated with compositions of the invention. Figures 3 and 8 are glyphs plotting cancer survival rates in animal models treated with or without compositions outside the scope of the present invention. Margin below % Procedural correction power (method) % formula % 1. Display of case 1988 Patent Application No. 60634 Name Danner-Farber Cancer Institute 4, Agent Address 105 Toranomon, Minato-ku, Tokyo - 8-10 Shizuka Toranobar Building Telephone 504-07216, Subject of amendment (1) Statement (2) Corporate certificate 7, Contents of amendment (11 Engraving of statement (no change in content) (2) As shown in Attachment 8, list of attached documents (1) 1 copy of engraving specification

Claims (1)

[Claims] 1. Structural formula ▲ Numerical formula, chemical formula, table, etc. ▼ (In the formula, X_1 and X_2 each independently represent O, S or S.
e, Z is an atom necessary to complete a saturated or aromatic condensed ring of 6-membered carbon ring atoms, and A is =CH-CH=
or ▲There are mathematical formulas, chemical formulas, tables, etc.▼ However, when X_2 is O, A is the latter group, and n is 0 or 1, and R_1, R
_2 and R_3 are each independently a methyl group or ethyl group, R_3 and R_4 are each independently a hydrogen atom or a halogen atom, and Q is a pharmaceutically acceptable anion) An anticancer composition comprising: